LOCUS       sequence-108759-        8235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding multiple cloning site, rybozyme and IRES-CAS9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8235)
  AUTHORS   Yoshioka S, Fujii W, Ogawa T, Sugiura K, Naito K
  TITLE     Development of a mono-promoter-driven CRISPR/Cas9 system in 
            mammalian cells.
  JOURNAL   Sci Rep. 2015 Dec 16;5:18341. doi: 10.1038/srep18341.
  PUBMED    26669567
REFERENCE   2  (bases 1 to 8235)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2015-12-16"; volume: "5:18341. doi"; pages: " 
            10.1038/srep18341"
COMMENT     Sequence Label: pMCS-rybozyme-IRES-CAS9
FEATURES             Location/Qualifiers
     source          1..8235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     226..245
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_RNA        301..376
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           381..448
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_feature    join(546..1120,1121..1123,1124..1132)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(713..730)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     940..959
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1136..1159
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1169..1189
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1214..5314
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1214..5314
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1214..5314
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5327..5347
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5371..5390)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5436..5491
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5490..5509)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5852..5868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5852..5868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5865..5887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5876..5892
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5900..5906,5907..5924,5925..5930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5945..5966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6025..6220
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6071..6206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6226..6360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6263..6282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6317..6336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6428..6445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6599..7187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6679..6698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7358..8149,8150..8218))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7981..8000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8219..8235,1..88))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
       61 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga
      121 cgacgtcacc ggtgtacagc tagcgcggcc gcgtataccc gcgggcatgc tctagagcct
      181 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
      241 ttgtgctgtc tcatcatttt ggcaaagaat tcgattccgg agagacggga tcccgtctct
      301 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      361 ggcaccgagt cggtgctttt ggccggcatg gtcccagcct cctcgctggc gccggctggg
      421 caacatgctt cggcatggcg aatgggacaa tcactagtga attcactcct caggtgcagg
      481 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
      541 atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
      601 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
      661 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
      721 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
      781 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
      841 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
      901 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
      961 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     1021 cggtacacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     1081 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggatta
     1141 caaggatgac gacgataaga tcatggcccc aaagaagaag cggaaggtcg gtatccacgg
     1201 agtcccagca gccgacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg
     1261 ctgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
     1321 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
     1381 gaccgccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
     1441 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
     1501 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
     1561 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
     1621 tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc
     1681 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
     1741 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     1801 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     1861 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
     1921 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
     1981 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
     2041 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
     2101 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
     2161 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
     2221 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
     2281 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
     2341 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
     2401 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
     2461 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
     2521 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
     2581 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
     2641 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
     2701 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
     2761 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
     2821 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
     2881 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
     2941 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
     3001 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
     3061 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
     3121 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
     3181 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
     3241 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
     3301 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
     3361 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
     3421 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
     3481 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
     3541 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
     3601 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
     3661 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
     3721 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
     3781 gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa
     3841 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
     3901 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
     3961 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
     4021 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
     4081 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
     4141 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
     4201 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
     4261 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
     4321 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
     4381 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
     4441 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa
     4501 gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga
     4561 caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc
     4621 tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact
     4681 caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa
     4741 ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa
     4801 gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc
     4861 gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta
     4921 tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct
     4981 gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc
     5041 caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca
     5101 cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa
     5161 cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac
     5221 ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga
     5281 aacaagaatc gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag
     5341 gaaggtgtga atcgatggta caagtaagga tctttttccc tctgccaaaa attatgggga
     5401 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     5461 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     5521 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     5581 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5641 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5701 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     5761 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     5821 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5881 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5941 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6001 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     6061 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6121 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6181 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     6241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     6361 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7021 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7501 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8221 tcttcctttt tcaat
//
